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R&D Systems recombinant murine il 13
<t>IL-13</t> receptor expression and TGF-β 1 production after CT-26 injection. ( A ) IL-13Rα 1 and IL-13Rα 2 expression during the CT-26 lung tumor model in BALB/c mice. IL-13Rα 1 mRNA expression was determined by RT-PCR of RNA extracted
Recombinant Murine Il 13, supplied by R&D Systems, used in various techniques. Bioz Stars score: 93/100, based on 13 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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1) Product Images from "Restoration of Tumor Immune Surveillance via Targeting of IL-13Receptor-?2"

Article Title: Restoration of Tumor Immune Surveillance via Targeting of IL-13Receptor-?2

Journal:

doi: 10.1158/0008-5472.CAN-07-5301

IL-13 receptor expression and TGF-β 1 production after CT-26 injection. ( A ) IL-13Rα 1 and IL-13Rα 2 expression during the CT-26 lung tumor model in BALB/c mice. IL-13Rα 1 mRNA expression was determined by RT-PCR of RNA extracted
Figure Legend Snippet: IL-13 receptor expression and TGF-β 1 production after CT-26 injection. ( A ) IL-13Rα 1 and IL-13Rα 2 expression during the CT-26 lung tumor model in BALB/c mice. IL-13Rα 1 mRNA expression was determined by RT-PCR of RNA extracted

Techniques Used: Expressing, Injection, Mouse Assay, Reverse Transcription Polymerase Chain Reaction

2) Product Images from "Homeostatic control of conjunctival mucosal goblet cells by NKT-derived IL-13"

Article Title: Homeostatic control of conjunctival mucosal goblet cells by NKT-derived IL-13

Journal: Mucosal immunology

doi: 10.1038/mi.2010.82

Gene expression analysis of isolated NK populations. Relative fold expression in natural killer (NK)/natural killer T (NKT)-positive and -negative populations. mRNA transcript levels of interleukin 13 (IL-13), interferon-γ (IFN-γ), IL-17A, IL-17F, IL-22, IL-21, and IL-4 in freshly isolated NK positive (NK +) and negative (NK −) cells isolated cells from nonstressed (NS) and desiccating-stressed (DS) ocular surface (OS) for 5 (DS5) or 10 days (DS10) and normal spleen. *** P
Figure Legend Snippet: Gene expression analysis of isolated NK populations. Relative fold expression in natural killer (NK)/natural killer T (NKT)-positive and -negative populations. mRNA transcript levels of interleukin 13 (IL-13), interferon-γ (IFN-γ), IL-17A, IL-17F, IL-22, IL-21, and IL-4 in freshly isolated NK positive (NK +) and negative (NK −) cells isolated cells from nonstressed (NS) and desiccating-stressed (DS) ocular surface (OS) for 5 (DS5) or 10 days (DS10) and normal spleen. *** P

Techniques Used: Expressing, Isolation

NK and NKT cells produce IL-13. ( a ) Mean±s.d. of interleukin 13 (IL-13) enzyme-linked immunosorbent spots (ELISPOTs) after γδ + isolation from nonstressed (NS) ocular surface (OS) and NS spleen, showing the three isolated populations (αβ +, γδ +, and B220 and CD11b + cells). ( b ) Mean±s. d. of IL-13 ELISPOTs after natural killer (NK) isolation from NS and desiccated-stressed (DS) OS for 10 days (DS10) and NS spleen, showing the two isolated populations, NK positive ( + ) and NK negative ( − ). ( c ) Histogram of flow cytometry analysis of total splenocytes stained with NK1.1-phycoerythrin (PE)-conjugated antibody (gray line) or T-cell receptor αβ-antigen-presenting cell (TCRαβ-APC) and isotype control antibody (black line). The number over line indicates percentage of positive cells over the isotype control antibody. ( d, e ) Flow cytometry analysis. Lymphocytes were gated based on characteristic light-scatter properties, and single lymphocytes were gated based on forward scatter height vs. forward scatter area (FSC-A). The numbers in the quadrants or over line indicate the percentage of cells. ( d ) Histogram of flow cytometry analysis of NK-positive cells stained with NK1.1-PE-conjugated antibody (gray line) or isotype control antibody (black line). The number over line indicates percentage of positive cells over the isotype control antibody. ( e ) Flow cytometry analysis of freshly isolated NK cells dual stained with NK1.1-PE + TCRαβ in all populations (all cells=no fraction, NK +, and NK − cells) from NS spleens. Percentage of double-positive cells is indicated in top right quadrant. ( f ) Mean±s.d. of IL-13 ELISPOTs after CD45 isolation from NS OS.
Figure Legend Snippet: NK and NKT cells produce IL-13. ( a ) Mean±s.d. of interleukin 13 (IL-13) enzyme-linked immunosorbent spots (ELISPOTs) after γδ + isolation from nonstressed (NS) ocular surface (OS) and NS spleen, showing the three isolated populations (αβ +, γδ +, and B220 and CD11b + cells). ( b ) Mean±s. d. of IL-13 ELISPOTs after natural killer (NK) isolation from NS and desiccated-stressed (DS) OS for 10 days (DS10) and NS spleen, showing the two isolated populations, NK positive ( + ) and NK negative ( − ). ( c ) Histogram of flow cytometry analysis of total splenocytes stained with NK1.1-phycoerythrin (PE)-conjugated antibody (gray line) or T-cell receptor αβ-antigen-presenting cell (TCRαβ-APC) and isotype control antibody (black line). The number over line indicates percentage of positive cells over the isotype control antibody. ( d, e ) Flow cytometry analysis. Lymphocytes were gated based on characteristic light-scatter properties, and single lymphocytes were gated based on forward scatter height vs. forward scatter area (FSC-A). The numbers in the quadrants or over line indicate the percentage of cells. ( d ) Histogram of flow cytometry analysis of NK-positive cells stained with NK1.1-PE-conjugated antibody (gray line) or isotype control antibody (black line). The number over line indicates percentage of positive cells over the isotype control antibody. ( e ) Flow cytometry analysis of freshly isolated NK cells dual stained with NK1.1-PE + TCRαβ in all populations (all cells=no fraction, NK +, and NK − cells) from NS spleens. Percentage of double-positive cells is indicated in top right quadrant. ( f ) Mean±s.d. of IL-13 ELISPOTs after CD45 isolation from NS OS.

Techniques Used: Isolation, Flow Cytometry, Cytometry, Staining

Effects of CsA treatment on NK and NKT cells. ( a ) Mean±s.d. of conjunctival goblet cell (GC) density in nonstressed (NS) C57BL/6 control mice and mice subjected to desiccating stress for 5 or 10 days (DS5 and DS10, respectively) or DS5 treated with Cyclosporine A (DS5 + CsA) or DS5 treated with CsA vehicle (DS5 + Veh). ( b ) mRNA transcript levels of MUC5AC in conjunctiva of NS C57BL/6 control mice and mice subjected to DS5 and DS10, respectively. ( c ) Mean±.d. of protein concentrations of MUC5AC in conjunctiva of NS C57BL/6 control mice and mice subjected to DS5 and DS10, respectively. ( d, e ) Representative flow cytometry analysis of freshly isolated ocular surface cells stained with pan-NK marker DX5 –fluorescein isothiocyanate (FITC)-conjugated antibody in NS C57BL/6 controls mice and mice subjected to DS5 and DS10, respectively ( d ) or DS5 treated with CsA (DS5 + CsA) or DS5 treated with CsA vehicle (DS5 + Veh) ( e ). Lymphocytes were gated based on characteristic light-scatter properties, and single lymphocytes were gated based on forward scatter height vs. forward scatter area (FSC-A). Numbers in the quadrants indicate the percentage of cells. ( f ) Mean±s.d. of the percentage of DX5+ cells evaluated by flow cytometry in three independent experiments. ( g ) Percentage change in the level of expression of interferon-γ (IFN-γ), IL-13, IL-17A, IL-21, and IL-22 mRNA transcripts in CsA-treated NK + and NK − cell populations, compared with vehicle. *** P
Figure Legend Snippet: Effects of CsA treatment on NK and NKT cells. ( a ) Mean±s.d. of conjunctival goblet cell (GC) density in nonstressed (NS) C57BL/6 control mice and mice subjected to desiccating stress for 5 or 10 days (DS5 and DS10, respectively) or DS5 treated with Cyclosporine A (DS5 + CsA) or DS5 treated with CsA vehicle (DS5 + Veh). ( b ) mRNA transcript levels of MUC5AC in conjunctiva of NS C57BL/6 control mice and mice subjected to DS5 and DS10, respectively. ( c ) Mean±.d. of protein concentrations of MUC5AC in conjunctiva of NS C57BL/6 control mice and mice subjected to DS5 and DS10, respectively. ( d, e ) Representative flow cytometry analysis of freshly isolated ocular surface cells stained with pan-NK marker DX5 –fluorescein isothiocyanate (FITC)-conjugated antibody in NS C57BL/6 controls mice and mice subjected to DS5 and DS10, respectively ( d ) or DS5 treated with CsA (DS5 + CsA) or DS5 treated with CsA vehicle (DS5 + Veh) ( e ). Lymphocytes were gated based on characteristic light-scatter properties, and single lymphocytes were gated based on forward scatter height vs. forward scatter area (FSC-A). Numbers in the quadrants indicate the percentage of cells. ( f ) Mean±s.d. of the percentage of DX5+ cells evaluated by flow cytometry in three independent experiments. ( g ) Percentage change in the level of expression of interferon-γ (IFN-γ), IL-13, IL-17A, IL-21, and IL-22 mRNA transcripts in CsA-treated NK + and NK − cell populations, compared with vehicle. *** P

Techniques Used: Mouse Assay, Flow Cytometry, Cytometry, Isolation, Staining, Marker, Expressing

IL-13 expression and goblet cell analysis. ( a–c ) Immunohistochemical staining for interleukin 13 (IL-13; red, in a) and IL-13 receptor α1 (IL-13Rα1; red, in b) and goat anti-rabbit secondary antibody alone ( c ) in conjunctiva sections of nonstressed (NS) control mice. Scale bar = 25 μm. ( d ) mRNA transcript levels of IL-13 and interferon-γ (IFN-γ) in conjunctiva in NS control mice and mice subjected to desiccating stress for 5 or 10 days (DS5 and DS10, respectively). ( e ) Ratio of IL-13/IFN-γ mRNA transcripts in conjunctiva. ( f ) Mean±s.d. of goblet cell (GC) density in NS C57BL/6 (B6) control mice and mice subjected to DS5 and mice after subconjunctival injection of BSA (+ BSA) or IL-13 (+ IL13). ( g ) Mean±s.d. of GC density in NS C57BL/6 (B6) controls mice and mice subjected to DS5 and IL-13 knockout (KO) mice. ( h ) Mean±s.d. of GC density in NS Bal/c (BC) control mice and mice subjected to DS5 and DS10 and signal transducer and activator of transcription 6 knockout (STAT6KO) at baseline and after DS5. ( i ) Mean±s. d. of GC density in NS C57BL/6 mice (B6) that received systemic injection of neutralizing antibody (NK1.1) to natural killer (NK) cells (αNK) or isotype control (IC) antibody before NS and after DS5. ( j ) Mean±s.d. of GC density in NS C57BL/6 (B6) control and RAG1KO mice. ( k ) Mean±s.d. of GC density in NS Bal/c (BC) control mice and CD1dKO at baseline, NS, and after DS5. A separate group NS CD1dKO mice received systemic injection of depleting antibody (NK1.1) to NK cells (αNK) or IC antibody.
Figure Legend Snippet: IL-13 expression and goblet cell analysis. ( a–c ) Immunohistochemical staining for interleukin 13 (IL-13; red, in a) and IL-13 receptor α1 (IL-13Rα1; red, in b) and goat anti-rabbit secondary antibody alone ( c ) in conjunctiva sections of nonstressed (NS) control mice. Scale bar = 25 μm. ( d ) mRNA transcript levels of IL-13 and interferon-γ (IFN-γ) in conjunctiva in NS control mice and mice subjected to desiccating stress for 5 or 10 days (DS5 and DS10, respectively). ( e ) Ratio of IL-13/IFN-γ mRNA transcripts in conjunctiva. ( f ) Mean±s.d. of goblet cell (GC) density in NS C57BL/6 (B6) control mice and mice subjected to DS5 and mice after subconjunctival injection of BSA (+ BSA) or IL-13 (+ IL13). ( g ) Mean±s.d. of GC density in NS C57BL/6 (B6) controls mice and mice subjected to DS5 and IL-13 knockout (KO) mice. ( h ) Mean±s.d. of GC density in NS Bal/c (BC) control mice and mice subjected to DS5 and DS10 and signal transducer and activator of transcription 6 knockout (STAT6KO) at baseline and after DS5. ( i ) Mean±s. d. of GC density in NS C57BL/6 mice (B6) that received systemic injection of neutralizing antibody (NK1.1) to natural killer (NK) cells (αNK) or isotype control (IC) antibody before NS and after DS5. ( j ) Mean±s.d. of GC density in NS C57BL/6 (B6) control and RAG1KO mice. ( k ) Mean±s.d. of GC density in NS Bal/c (BC) control mice and CD1dKO at baseline, NS, and after DS5. A separate group NS CD1dKO mice received systemic injection of depleting antibody (NK1.1) to NK cells (αNK) or IC antibody.

Techniques Used: Expressing, Immunohistochemistry, Staining, Mouse Assay, Injection, Knock-Out

3) Product Images from "IL-13 PROTECTS MOUSE INTESTINE FROM ISCHEMIA AND REPERFUSION INJURY THROUGH REGULATION OF INNATE AND ADAPTIVE IMMUNITY"

Article Title: IL-13 PROTECTS MOUSE INTESTINE FROM ISCHEMIA AND REPERFUSION INJURY THROUGH REGULATION OF INNATE AND ADAPTIVE IMMUNITY

Journal: Transplantation

doi: 10.1097/TP.0b013e31820c861a

The effects of intestinal I/R injury on mouse survival. C57BL6 wild-type (WT) mice were either treated with rIL-13, anti-IL-13, anti-IL-13+rIL-13, or saline (control). 7 days survival in these groups was 100%, 0%, 0%, and 50%, respectively (n = 6 mice/group).
Figure Legend Snippet: The effects of intestinal I/R injury on mouse survival. C57BL6 wild-type (WT) mice were either treated with rIL-13, anti-IL-13, anti-IL-13+rIL-13, or saline (control). 7 days survival in these groups was 100%, 0%, 0%, and 50%, respectively (n = 6 mice/group).

Techniques Used: Mouse Assay

Competitive template RT-PCR assisted expression of mRNA coding for cytokine IL-2/IFN-γ and IL-4/IL-13, and chemokine IP-10/MCP-1 in ischemic intestines at 4 h and 24 h after 100 minutes warm ischemia. Note: The expression of IL-2, IFN-γ,
Figure Legend Snippet: Competitive template RT-PCR assisted expression of mRNA coding for cytokine IL-2/IFN-γ and IL-4/IL-13, and chemokine IP-10/MCP-1 in ischemic intestines at 4 h and 24 h after 100 minutes warm ischemia. Note: The expression of IL-2, IFN-γ,

Techniques Used: Reverse Transcription Polymerase Chain Reaction, Expressing

4) Product Images from "Bone marrow myeloid-derived suppressor cells (MDSCs) inhibit graft-versus-host disease (GVHD) via an arginase-1-dependent mechanism that is up-regulated by interleukin-13"

Article Title: Bone marrow myeloid-derived suppressor cells (MDSCs) inhibit graft-versus-host disease (GVHD) via an arginase-1-dependent mechanism that is up-regulated by interleukin-13

Journal: Blood

doi: 10.1182/blood-2010-06-287839

MDSC-IL-13 negatively impact proliferation, activation, and effector function of donor T cells . Lethally irradiated BALB/c mice were transplanted with 15 × 10 6 CFSE-labeled CD25-depleted B6 Ly5.1-expressing T cells alone or with 10 × 10
Figure Legend Snippet: MDSC-IL-13 negatively impact proliferation, activation, and effector function of donor T cells . Lethally irradiated BALB/c mice were transplanted with 15 × 10 6 CFSE-labeled CD25-depleted B6 Ly5.1-expressing T cells alone or with 10 × 10

Techniques Used: Activation Assay, Irradiation, Mouse Assay, Labeling, Expressing

PEG-arg1 has similar protective effects as MDSC IL-13-tx . Lethally irradiated BALB/c mice were given 10 × 10 6 B6 BM cells alone or with 2 × 10 6 CD25-depleted T cells. In addition to this, cohorts were also given 6 × 10 6 MDSC IL-13
Figure Legend Snippet: PEG-arg1 has similar protective effects as MDSC IL-13-tx . Lethally irradiated BALB/c mice were given 10 × 10 6 B6 BM cells alone or with 2 × 10 6 CD25-depleted T cells. In addition to this, cohorts were also given 6 × 10 6 MDSC IL-13

Techniques Used: Irradiation, Mouse Assay

MDSCs inhibit T-cell alloresponses through the expression of arginase-1 . (A) RT-PCR showing that MDSCs express arginase-1 and iNOS and up-regulate arginase-1 on stimulation with IL-13. Hypoxanthine phosphoribosyl transferase shown as endogenous control.
Figure Legend Snippet: MDSCs inhibit T-cell alloresponses through the expression of arginase-1 . (A) RT-PCR showing that MDSCs express arginase-1 and iNOS and up-regulate arginase-1 on stimulation with IL-13. Hypoxanthine phosphoribosyl transferase shown as endogenous control.

Techniques Used: Expressing, Reverse Transcription Polymerase Chain Reaction

MDSC-IL-13 migrate to sites of allopriming in a GVHD setting . (A) Fluorescence-activated cell sorter analysis of pretransplantation MDSC-IL-13 was gated on Gr1 + CD11b + and analyzed for the surface expression of molecules associated with homing and recruitment.
Figure Legend Snippet: MDSC-IL-13 migrate to sites of allopriming in a GVHD setting . (A) Fluorescence-activated cell sorter analysis of pretransplantation MDSC-IL-13 was gated on Gr1 + CD11b + and analyzed for the surface expression of molecules associated with homing and recruitment.

Techniques Used: Fluorescence, Expressing

Cultured MDSC-IL-13 enhance GVHD survival in an arginase-1-dependent fashion . Lethally irradiated BALB/c recipients were given 10 7 B6 BM cells plus 2 × 10 6 purified CD25-depleted T cells plus either 2 × 10 6 MDSC-IL-13 or 6 × 10
Figure Legend Snippet: Cultured MDSC-IL-13 enhance GVHD survival in an arginase-1-dependent fashion . Lethally irradiated BALB/c recipients were given 10 7 B6 BM cells plus 2 × 10 6 purified CD25-depleted T cells plus either 2 × 10 6 MDSC-IL-13 or 6 × 10

Techniques Used: Cell Culture, Irradiation, Purification

5) Product Images from "Innate lymphoid cells promote lung tissue homeostasis following acute influenza virus infection"

Article Title: Innate lymphoid cells promote lung tissue homeostasis following acute influenza virus infection

Journal: Nature Immunology

doi: 10.1031/ni.2131

ILCs in the lung resemble nuocytes and NHCs in phenotype and cytokine profile ( a ) Identification of lung ILCs in C57BL/6 wild-type (WT) and Rag1 −/− mice by flow cytometry as CD90 + CD25 + CD127 + lineage (Lin) negative cells lacking expression of the follow markers: (CD3, CD5, NK1.1, CD27, TCRβ antibodies on the y-axis, B220, CD11b, CD11c antibodies on the x -axis). ( b ) Expression of cell surface markers on Lin − CD90 + CD25 + lung ILCs in C57BL/6 WT (solid black line) and Rag1 −/− mice (dashed black line) compared to isotype controls (gray shaded). ( a-b ) Data is representative of more than three experiments, n = at least 4 WT or Rag1 −/− mice. ( c ) Absolute number of CD90 + CD25 + ILCs in naïve WT or Rag1 −/− lung. d ) Flow cytometry plots of pre-sort and post-sort purity of CD90 + CD25 + T1-ST2 + WT lung ILCs, gated on live, lineage negative cells. ( e ) IL-5 and IL-13 cytokine secretion from flow sorted CD90 + CD25 + T1-ST2 + lung ILCs cultured with IL-2 + IL-7 alone or in combination with IL-33 for four days, measured by ELISA. Data is representative of three independent experiments, n = 3 replicates, each replicate consisting of ILCs sorted from 5 pooled lungs. ( f ) Intracellular cytokine staining for IL-22 and IL-17A in Lin − CD90 + CD25 + lung ILCs or spleen CD90 + CD4 + LTi cells from WT mice, stimulated with 50 ng/ml rIL-23 (12 h) + PMA and Ionomycin (4 h). ( g ) IL-17A cytokine secretion from flow sorted CD90 + CD25 + T1-ST2 + lung ILCs or spleen CD90 + CD4 + LTi cells cultured with IL-2 + IL-7 alone or in combination with IL-23 for four days, measured by ELISA. Data is representative of two independent experiments, n = 3 replicates, each replicate consisting of ILCs sorted from 3 pooled lungs or spleens. ( h ) Intracellular cytokine staining in Lin − CD90 + CD25 + lung ILCs from WT mice treated with 500 ng rIL-33 for 7 days in vivo and stimulated with PMA + Ionomycin (4 hours) ex vivo . ( f,h ) Data is representative of 2 or more experiments, n = 3–4 mice.
Figure Legend Snippet: ILCs in the lung resemble nuocytes and NHCs in phenotype and cytokine profile ( a ) Identification of lung ILCs in C57BL/6 wild-type (WT) and Rag1 −/− mice by flow cytometry as CD90 + CD25 + CD127 + lineage (Lin) negative cells lacking expression of the follow markers: (CD3, CD5, NK1.1, CD27, TCRβ antibodies on the y-axis, B220, CD11b, CD11c antibodies on the x -axis). ( b ) Expression of cell surface markers on Lin − CD90 + CD25 + lung ILCs in C57BL/6 WT (solid black line) and Rag1 −/− mice (dashed black line) compared to isotype controls (gray shaded). ( a-b ) Data is representative of more than three experiments, n = at least 4 WT or Rag1 −/− mice. ( c ) Absolute number of CD90 + CD25 + ILCs in naïve WT or Rag1 −/− lung. d ) Flow cytometry plots of pre-sort and post-sort purity of CD90 + CD25 + T1-ST2 + WT lung ILCs, gated on live, lineage negative cells. ( e ) IL-5 and IL-13 cytokine secretion from flow sorted CD90 + CD25 + T1-ST2 + lung ILCs cultured with IL-2 + IL-7 alone or in combination with IL-33 for four days, measured by ELISA. Data is representative of three independent experiments, n = 3 replicates, each replicate consisting of ILCs sorted from 5 pooled lungs. ( f ) Intracellular cytokine staining for IL-22 and IL-17A in Lin − CD90 + CD25 + lung ILCs or spleen CD90 + CD4 + LTi cells from WT mice, stimulated with 50 ng/ml rIL-23 (12 h) + PMA and Ionomycin (4 h). ( g ) IL-17A cytokine secretion from flow sorted CD90 + CD25 + T1-ST2 + lung ILCs or spleen CD90 + CD4 + LTi cells cultured with IL-2 + IL-7 alone or in combination with IL-23 for four days, measured by ELISA. Data is representative of two independent experiments, n = 3 replicates, each replicate consisting of ILCs sorted from 3 pooled lungs or spleens. ( h ) Intracellular cytokine staining in Lin − CD90 + CD25 + lung ILCs from WT mice treated with 500 ng rIL-33 for 7 days in vivo and stimulated with PMA + Ionomycin (4 hours) ex vivo . ( f,h ) Data is representative of 2 or more experiments, n = 3–4 mice.

Techniques Used: Mouse Assay, Flow Cytometry, Cytometry, Expressing, Cell Culture, Enzyme-linked Immunosorbent Assay, Staining, In Vivo, Ex Vivo

6) Product Images from "IL-13 Activates STAT6 and Inhibits Liver Injury Induced by Ischemia/Reperfusion"

Article Title: IL-13 Activates STAT6 and Inhibits Liver Injury Induced by Ischemia/Reperfusion

Journal: The American Journal of Pathology

doi:

Effects of IL-13 on mRNA expression for TNFα ( A ) and MIP-2 ( B ) as well as serum levels of TNFα ( C ) and MIP-2 ( D ) protein. Liver RNA extracts and serum samples were obtained from sham control mice and mice undergoing ischemia and 4 hours of reperfusion treated with saline (I/R) or IL-13 (I/R + IL-13). Liver RNA extracts were assessed by RT-PCR ( A and B ). Results are representative of three independent experiments. Serum samples were analyzed by ELISA ( C and D ). Values represent mean ± SE with n = 10 per group.
Figure Legend Snippet: Effects of IL-13 on mRNA expression for TNFα ( A ) and MIP-2 ( B ) as well as serum levels of TNFα ( C ) and MIP-2 ( D ) protein. Liver RNA extracts and serum samples were obtained from sham control mice and mice undergoing ischemia and 4 hours of reperfusion treated with saline (I/R) or IL-13 (I/R + IL-13). Liver RNA extracts were assessed by RT-PCR ( A and B ). Results are representative of three independent experiments. Serum samples were analyzed by ELISA ( C and D ). Values represent mean ± SE with n = 10 per group.

Techniques Used: Expressing, Mouse Assay, Reverse Transcription Polymerase Chain Reaction, Enzyme-linked Immunosorbent Assay

Effects of IL-13 on activation of NFκB and STAT6 in liver during hepatic ischemia/reperfusion (I/R) injury. A : EMSA analysis of NFκB and STAT6 in whole liver nuclear extracts from sham controls (I/R time: 0), ischemia and 1 hour reperfusion, and ischemia and 4 hours reperfusion in the presence or absence of IL-13. B : Competition and supershift assays were performed to verify the specificity of the STAT6 consensus oligonucleotide probe. Whole liver extracts obtained from mice treated with IL-13 and undergoing ischemia and 1 hour of reperfusion were incubated with 32 P-labeled STAT6 oligonucleotide in the absence (none) or presence of unlabeled (cold) NFκB oligonucleotide, unlabeled (cold) STAT6 oligonucleotide, or antibodies to STAT4 or STAT6.
Figure Legend Snippet: Effects of IL-13 on activation of NFκB and STAT6 in liver during hepatic ischemia/reperfusion (I/R) injury. A : EMSA analysis of NFκB and STAT6 in whole liver nuclear extracts from sham controls (I/R time: 0), ischemia and 1 hour reperfusion, and ischemia and 4 hours reperfusion in the presence or absence of IL-13. B : Competition and supershift assays were performed to verify the specificity of the STAT6 consensus oligonucleotide probe. Whole liver extracts obtained from mice treated with IL-13 and undergoing ischemia and 1 hour of reperfusion were incubated with 32 P-labeled STAT6 oligonucleotide in the absence (none) or presence of unlabeled (cold) NFκB oligonucleotide, unlabeled (cold) STAT6 oligonucleotide, or antibodies to STAT4 or STAT6.

Techniques Used: Activation Assay, Mouse Assay, Incubation, Labeling

Effects of IL-13 on liver neutrophil accumulation ( A and B ), hepatocellular injury ( C ), and liver edema ( D ) in sham control mice and mice undergoing ischemia and 4 hours of reperfusion treated with saline (I/R) or IL-13 (I/R + IL-13). Liver neutrophil accumulation was determined by measuring liver myeloperoxidase (MPO) content ( A ), as well as by liver tissue neutrophil morphometrics (PMN/high power field; B ). Hepatocellular injury was assessed by determining serum levels of alanine aminotransferase (ALT). Liver edema was quantitated by liver wet-to-dry weight ratios. Values represent mean ± SE with n = 10 per group.
Figure Legend Snippet: Effects of IL-13 on liver neutrophil accumulation ( A and B ), hepatocellular injury ( C ), and liver edema ( D ) in sham control mice and mice undergoing ischemia and 4 hours of reperfusion treated with saline (I/R) or IL-13 (I/R + IL-13). Liver neutrophil accumulation was determined by measuring liver myeloperoxidase (MPO) content ( A ), as well as by liver tissue neutrophil morphometrics (PMN/high power field; B ). Hepatocellular injury was assessed by determining serum levels of alanine aminotransferase (ALT). Liver edema was quantitated by liver wet-to-dry weight ratios. Values represent mean ± SE with n = 10 per group.

Techniques Used: Mouse Assay

7) Product Images from "Inactivation of paracellular cation-selective claudin-2 channels attenuates immune-mediated experimental colitis in mice"

Article Title: Inactivation of paracellular cation-selective claudin-2 channels attenuates immune-mediated experimental colitis in mice

Journal: The Journal of Clinical Investigation

doi: 10.1172/JCI138697

Claudin-2 is necessary and sufficient for IL-13–induced changes in pore pathway permeability. ( A ) Colonic histopathology of Cldn2 +/+ or Cldn2 –/– mice was not affected by injection with vehicle or IL-13. ( B ) IL-13 increases claudin-2 (CLDN2, green) protein expression in proximal colonic epithelial cells of Cldn2 +/+ , but not Cldn2 –/– , mice. Nuclei (blue) are shown for reference. ( C ) Immunoblots of isolated colonic epithelia from Cldn2 +/+ and Cldn2 –/– mice treated with vehicle or IL-13. Claudin-2, claudin-4 (CLDN4), occludin (OCLN), E-cadherin (ECAD), and β-actin are shown. ( D ) Densitometry of immunoblots, as in C . n = 3–4 per condition. ANOVA with Bonferroni’s correction. ( E and F ) Proximal colonic mucosae from Cldn2 +/+ ( E ) and Cldn2 –/– ( F ) mice treated with vehicle (circles) or IL-13 (squares) were mounted in Ussing chambers for analysis of paracellular permeability. Bi-ionic potential measurements were used to determine the permeabilities of Na + and 5 larger cations (methylamine, ethylamine, tetramethylammonium, tetraethylammonium, and N -methyl-D-glucamine). IL-13 increased permeability of Na + , methylamine, and ethylamine, but not larger cations, in Cldn2 +/+ mice. IL-13 did not affect Na + , methylamine, or ethylamine permeability in Cldn2 –/– mice. n = 8 and 9 for Cldn2 +/+ mice without or with IL-13 treatment, respectively, and n = 5 and 9 for Cldn2 –/– mice without or with IL-13 treatment, respectively. Data compiled from 3 independent experiments. Two-tailed t test. ( G ) Claudin-2 (green) expression in Cldn2 +/+ and Cldn2 Tg mice. ( H ) Representative immunoblots of isolated colonic epithelia from Cldn2 +/+ and Cldn2 Tg mice. ( I ) Densitometry of immunoblots of isolated colonic epithelia from Cldn2 +/+ and Cldn2 Tg mice, as in H . n = 3–4 per condition. Two-tailed t test. ( J ) Ussing chamber analysis (as in E ) of proximal colonic mucosae from Cldn2 +/+ and Cldn2 Tg mice. Claudin-2 overexpression selectively increased Na + , methylamine, and ethylamine permeability. n = 11 Cldn2 +/+ , 10 Cldn2 Tg . Data compiled from 3 independent experiments. Two-tailed t test. * P
Figure Legend Snippet: Claudin-2 is necessary and sufficient for IL-13–induced changes in pore pathway permeability. ( A ) Colonic histopathology of Cldn2 +/+ or Cldn2 –/– mice was not affected by injection with vehicle or IL-13. ( B ) IL-13 increases claudin-2 (CLDN2, green) protein expression in proximal colonic epithelial cells of Cldn2 +/+ , but not Cldn2 –/– , mice. Nuclei (blue) are shown for reference. ( C ) Immunoblots of isolated colonic epithelia from Cldn2 +/+ and Cldn2 –/– mice treated with vehicle or IL-13. Claudin-2, claudin-4 (CLDN4), occludin (OCLN), E-cadherin (ECAD), and β-actin are shown. ( D ) Densitometry of immunoblots, as in C . n = 3–4 per condition. ANOVA with Bonferroni’s correction. ( E and F ) Proximal colonic mucosae from Cldn2 +/+ ( E ) and Cldn2 –/– ( F ) mice treated with vehicle (circles) or IL-13 (squares) were mounted in Ussing chambers for analysis of paracellular permeability. Bi-ionic potential measurements were used to determine the permeabilities of Na + and 5 larger cations (methylamine, ethylamine, tetramethylammonium, tetraethylammonium, and N -methyl-D-glucamine). IL-13 increased permeability of Na + , methylamine, and ethylamine, but not larger cations, in Cldn2 +/+ mice. IL-13 did not affect Na + , methylamine, or ethylamine permeability in Cldn2 –/– mice. n = 8 and 9 for Cldn2 +/+ mice without or with IL-13 treatment, respectively, and n = 5 and 9 for Cldn2 –/– mice without or with IL-13 treatment, respectively. Data compiled from 3 independent experiments. Two-tailed t test. ( G ) Claudin-2 (green) expression in Cldn2 +/+ and Cldn2 Tg mice. ( H ) Representative immunoblots of isolated colonic epithelia from Cldn2 +/+ and Cldn2 Tg mice. ( I ) Densitometry of immunoblots of isolated colonic epithelia from Cldn2 +/+ and Cldn2 Tg mice, as in H . n = 3–4 per condition. Two-tailed t test. ( J ) Ussing chamber analysis (as in E ) of proximal colonic mucosae from Cldn2 +/+ and Cldn2 Tg mice. Claudin-2 overexpression selectively increased Na + , methylamine, and ethylamine permeability. n = 11 Cldn2 +/+ , 10 Cldn2 Tg . Data compiled from 3 independent experiments. Two-tailed t test. * P

Techniques Used: Permeability, Histopathology, Mouse Assay, Injection, Expressing, Western Blot, Isolation, Two Tailed Test, Over Expression

8) Product Images from "IL-13 signaling via IL-13R?2 triggers TGF-?1-dependent allograft fibrosis"

Article Title: IL-13 signaling via IL-13R?2 triggers TGF-?1-dependent allograft fibrosis

Journal: Transplantation Research

doi: 10.1186/2047-1440-2-16

Activation of IL-13/TGF-β 1 pathway in allogeneically transplanted grafts. (A) ELISA of supernants of cultured allograft-infiltrating cells detected significantly elevated IL-13 levels in allografts (day 60, P = 0.0031 and day 100, P = 0.0003)compared to syngrafts or FVB control hearts ( P = 0.0055 and P = 0.0042). (B) Immunohistochemistry showed significantly higher and over time increasing numbers of IL-13 + cells in FVB hearts transplanted into DBA/1 recipients (day 60, P = 0.0228; day 100, P = 0.0037) relative to syngeneic animals (day 60 versus day 100, P = 0.0083). (C) Representative immunohistochemical stainings showed more IL-13 + cells in allografts ( FVB into DBA/1) compared to controls (DBA/1 into DBA/1; day 100). (D) Western blot analysis revealed expression of IL-13Rα 2 only in allograft-infiltrating cells isolated from allogeneically transplanted hearts (FVB into DBA/1) in contrast to cells isolated from FVB controls or syngrafts (DBA/1 into DBA/1) without IL-13Rα 2 expression. (E) Measurement of TGF-β 1 by ELISA detected significantly elevated TGF-β 1 levels in cells isolated from DBA/1 mice grafted with FVB hearts at day 60 (88 ± 5 versus 46 ± 5 pg/mL; P = 0.0010) and at day 100 (133 ± 6 versus 42 ± 7 pg/mL; P
Figure Legend Snippet: Activation of IL-13/TGF-β 1 pathway in allogeneically transplanted grafts. (A) ELISA of supernants of cultured allograft-infiltrating cells detected significantly elevated IL-13 levels in allografts (day 60, P = 0.0031 and day 100, P = 0.0003)compared to syngrafts or FVB control hearts ( P = 0.0055 and P = 0.0042). (B) Immunohistochemistry showed significantly higher and over time increasing numbers of IL-13 + cells in FVB hearts transplanted into DBA/1 recipients (day 60, P = 0.0228; day 100, P = 0.0037) relative to syngeneic animals (day 60 versus day 100, P = 0.0083). (C) Representative immunohistochemical stainings showed more IL-13 + cells in allografts ( FVB into DBA/1) compared to controls (DBA/1 into DBA/1; day 100). (D) Western blot analysis revealed expression of IL-13Rα 2 only in allograft-infiltrating cells isolated from allogeneically transplanted hearts (FVB into DBA/1) in contrast to cells isolated from FVB controls or syngrafts (DBA/1 into DBA/1) without IL-13Rα 2 expression. (E) Measurement of TGF-β 1 by ELISA detected significantly elevated TGF-β 1 levels in cells isolated from DBA/1 mice grafted with FVB hearts at day 60 (88 ± 5 versus 46 ± 5 pg/mL; P = 0.0010) and at day 100 (133 ± 6 versus 42 ± 7 pg/mL; P

Techniques Used: Activation Assay, Enzyme-linked Immunosorbent Assay, Cell Culture, Immunohistochemistry, Western Blot, Expressing, Isolation, Mouse Assay

9) Product Images from "Inactivation of paracellular cation-selective claudin-2 channels attenuates immune-mediated experimental colitis in mice"

Article Title: Inactivation of paracellular cation-selective claudin-2 channels attenuates immune-mediated experimental colitis in mice

Journal: The Journal of Clinical Investigation

doi: 10.1172/JCI138697

Claudin-2 is necessary and sufficient for IL-13–induced changes in pore pathway permeability. ( A ) Colonic histopathology of Cldn2 +/+ or Cldn2 –/– mice was not affected by injection with vehicle or IL-13. ( B ) IL-13 increases claudin-2 (CLDN2, green) protein expression in proximal colonic epithelial cells of Cldn2 +/+ , but not Cldn2 –/– , mice. Nuclei (blue) are shown for reference. ( C ) Immunoblots of isolated colonic epithelia from Cldn2 +/+ and Cldn2 –/– mice treated with vehicle or IL-13. Claudin-2, claudin-4 (CLDN4), occludin (OCLN), E-cadherin (ECAD), and β-actin are shown. ( D ) Densitometry of immunoblots, as in C . n = 3–4 per condition. ANOVA with Bonferroni’s correction. ( E and F ) Proximal colonic mucosae from Cldn2 +/+ ( E ) and Cldn2 –/– ( F ) mice treated with vehicle (circles) or IL-13 (squares) were mounted in Ussing chambers for analysis of paracellular permeability. Bi-ionic potential measurements were used to determine the permeabilities of Na + and 5 larger cations (methylamine, ethylamine, tetramethylammonium, tetraethylammonium, and N -methyl-D-glucamine). IL-13 increased permeability of Na + , methylamine, and ethylamine, but not larger cations, in Cldn2 +/+ mice. IL-13 did not affect Na + , methylamine, or ethylamine permeability in Cldn2 –/– mice. n = 8 and 9 for Cldn2 +/+ mice without or with IL-13 treatment, respectively, and n = 5 and 9 for Cldn2 –/– mice without or with IL-13 treatment, respectively. Data compiled from 3 independent experiments. Two-tailed t test. ( G ) Claudin-2 (green) expression in Cldn2 +/+ and Cldn2 Tg mice. ( H ) Representative immunoblots of isolated colonic epithelia from Cldn2 +/+ and Cldn2 Tg mice. ( I ) Densitometry of immunoblots of isolated colonic epithelia from Cldn2 +/+ and Cldn2 Tg mice, as in H . n = 3–4 per condition. Two-tailed t test. ( J ) Ussing chamber analysis (as in E ) of proximal colonic mucosae from Cldn2 +/+ and Cldn2 Tg mice. Claudin-2 overexpression selectively increased Na + , methylamine, and ethylamine permeability. n = 11 Cldn2 +/+ , 10 Cldn2 Tg . Data compiled from 3 independent experiments. Two-tailed t test. * P
Figure Legend Snippet: Claudin-2 is necessary and sufficient for IL-13–induced changes in pore pathway permeability. ( A ) Colonic histopathology of Cldn2 +/+ or Cldn2 –/– mice was not affected by injection with vehicle or IL-13. ( B ) IL-13 increases claudin-2 (CLDN2, green) protein expression in proximal colonic epithelial cells of Cldn2 +/+ , but not Cldn2 –/– , mice. Nuclei (blue) are shown for reference. ( C ) Immunoblots of isolated colonic epithelia from Cldn2 +/+ and Cldn2 –/– mice treated with vehicle or IL-13. Claudin-2, claudin-4 (CLDN4), occludin (OCLN), E-cadherin (ECAD), and β-actin are shown. ( D ) Densitometry of immunoblots, as in C . n = 3–4 per condition. ANOVA with Bonferroni’s correction. ( E and F ) Proximal colonic mucosae from Cldn2 +/+ ( E ) and Cldn2 –/– ( F ) mice treated with vehicle (circles) or IL-13 (squares) were mounted in Ussing chambers for analysis of paracellular permeability. Bi-ionic potential measurements were used to determine the permeabilities of Na + and 5 larger cations (methylamine, ethylamine, tetramethylammonium, tetraethylammonium, and N -methyl-D-glucamine). IL-13 increased permeability of Na + , methylamine, and ethylamine, but not larger cations, in Cldn2 +/+ mice. IL-13 did not affect Na + , methylamine, or ethylamine permeability in Cldn2 –/– mice. n = 8 and 9 for Cldn2 +/+ mice without or with IL-13 treatment, respectively, and n = 5 and 9 for Cldn2 –/– mice without or with IL-13 treatment, respectively. Data compiled from 3 independent experiments. Two-tailed t test. ( G ) Claudin-2 (green) expression in Cldn2 +/+ and Cldn2 Tg mice. ( H ) Representative immunoblots of isolated colonic epithelia from Cldn2 +/+ and Cldn2 Tg mice. ( I ) Densitometry of immunoblots of isolated colonic epithelia from Cldn2 +/+ and Cldn2 Tg mice, as in H . n = 3–4 per condition. Two-tailed t test. ( J ) Ussing chamber analysis (as in E ) of proximal colonic mucosae from Cldn2 +/+ and Cldn2 Tg mice. Claudin-2 overexpression selectively increased Na + , methylamine, and ethylamine permeability. n = 11 Cldn2 +/+ , 10 Cldn2 Tg . Data compiled from 3 independent experiments. Two-tailed t test. * P

Techniques Used: Permeability, Histopathology, Mouse Assay, Injection, Expressing, Western Blot, Isolation, Two Tailed Test, Over Expression

10) Product Images from "Inhibition of the mitochondrial calcium uniporter prevents IL-13 and allergen-mediated airway epithelial apoptosis and loss of barrier function"

Article Title: Inhibition of the mitochondrial calcium uniporter prevents IL-13 and allergen-mediated airway epithelial apoptosis and loss of barrier function

Journal: Experimental cell research

doi: 10.1016/j.yexcr.2017.12.003

MCU inhibition blocks mitochondrial matrix Ca 2+ uptake in human airway epithelial cells. HAEC were infected with empty virus (Mt-GFP) or DN-MCU (both at 10 MOI) for 48 hrs. (A) Immunoblots for MCU in isolated mitochondrial fractions from Mt-GFP or DN-MCU-infected HAEC. COXIV: loading control for mitochondria. (B) Ca 2+ concentration by o-cresolphthalein assay in mitochondria. Data were normalized to total mitochondrial protein measured by BCA assay (n = 3 independent experiments). (C) Comparison of mitochondrial calcium levels in HAEC infected with control or DN-MCU virus. Arrow indicates calcium addition. Increasing mitochondrial calcium results in a rapid increase in the fluorescent signal but the subsequent decline in the fluorescent signal, representing mitochondrial calcium uptake, is only observed in control-infected cells. D) Mitochondrial matrix Ca 2+ uptake by Rhod-2 fluorescence. Arrow denotes when histamine was added. Peak mitochondrial matrix Ca 2+ concentration represented as Rhod-2 fluorescence intensity. (n = 4 independent experiments). (E) Mitochondrial matrix Ca 2+ uptake by Rhod-2 fluorescence. Arrow denotes when IL-13 was added. Peak mitochondrial matrix Ca 2+ uptake represented as Rhod-2 fluorescence intensity (n = 3 independent experiments). (F) Representative images and quantification of Rhod-2 fluorescence two minutes after addition of IL-13 (n = 3 independent experiments). All data are the means ± SEM. Student’s two-tailed t-test was used. * p
Figure Legend Snippet: MCU inhibition blocks mitochondrial matrix Ca 2+ uptake in human airway epithelial cells. HAEC were infected with empty virus (Mt-GFP) or DN-MCU (both at 10 MOI) for 48 hrs. (A) Immunoblots for MCU in isolated mitochondrial fractions from Mt-GFP or DN-MCU-infected HAEC. COXIV: loading control for mitochondria. (B) Ca 2+ concentration by o-cresolphthalein assay in mitochondria. Data were normalized to total mitochondrial protein measured by BCA assay (n = 3 independent experiments). (C) Comparison of mitochondrial calcium levels in HAEC infected with control or DN-MCU virus. Arrow indicates calcium addition. Increasing mitochondrial calcium results in a rapid increase in the fluorescent signal but the subsequent decline in the fluorescent signal, representing mitochondrial calcium uptake, is only observed in control-infected cells. D) Mitochondrial matrix Ca 2+ uptake by Rhod-2 fluorescence. Arrow denotes when histamine was added. Peak mitochondrial matrix Ca 2+ concentration represented as Rhod-2 fluorescence intensity. (n = 4 independent experiments). (E) Mitochondrial matrix Ca 2+ uptake by Rhod-2 fluorescence. Arrow denotes when IL-13 was added. Peak mitochondrial matrix Ca 2+ uptake represented as Rhod-2 fluorescence intensity (n = 3 independent experiments). (F) Representative images and quantification of Rhod-2 fluorescence two minutes after addition of IL-13 (n = 3 independent experiments). All data are the means ± SEM. Student’s two-tailed t-test was used. * p

Techniques Used: Inhibition, Infection, Western Blot, Isolation, Concentration Assay, BIA-KA, Fluorescence, Two Tailed Test

MCU inhibition in HAEC decreases IL-13-mediated conductance and epithelial monolayer permeability. (A, B) HAEC in Transwell inserts were grown to confluence. Fresh medium was added every 2 days until cells excluded basolateral media. Cells were then infected with Mt-GFP or DN-MCU (10 MOI) for 48 hrs and TEER (ohm × cm 2 ) was monitored after IL-13 treatment (72 hr, 10 ng/ml). Conductance was calculated based on TEER values. (C) Epithelial permeability was assessed by the amount of TRITC-dextran (1 mg/ml) detected in the media of the basolateral compartment of the Transwell and represented as fold-change over GFP-control cells. (D) Representative immunofluorescent images for ZO-1 (green) in HAEC expressing Mt-GFP or DN-MCU after 72 hr treatment with IL-13 or control (63x). Nuclei TO-PRO (blue), negative staining control without primary antibody for Mt-GFP or DN-MCU. Scale bar 100μm. All data are means ± SEM for 3 independent experiments (n = 3 samples/treatment group). Analysis used one-way ANOVA with Tukey post hoc test. * p
Figure Legend Snippet: MCU inhibition in HAEC decreases IL-13-mediated conductance and epithelial monolayer permeability. (A, B) HAEC in Transwell inserts were grown to confluence. Fresh medium was added every 2 days until cells excluded basolateral media. Cells were then infected with Mt-GFP or DN-MCU (10 MOI) for 48 hrs and TEER (ohm × cm 2 ) was monitored after IL-13 treatment (72 hr, 10 ng/ml). Conductance was calculated based on TEER values. (C) Epithelial permeability was assessed by the amount of TRITC-dextran (1 mg/ml) detected in the media of the basolateral compartment of the Transwell and represented as fold-change over GFP-control cells. (D) Representative immunofluorescent images for ZO-1 (green) in HAEC expressing Mt-GFP or DN-MCU after 72 hr treatment with IL-13 or control (63x). Nuclei TO-PRO (blue), negative staining control without primary antibody for Mt-GFP or DN-MCU. Scale bar 100μm. All data are means ± SEM for 3 independent experiments (n = 3 samples/treatment group). Analysis used one-way ANOVA with Tukey post hoc test. * p

Techniques Used: Inhibition, Permeability, Infection, Expressing, Negative Staining

Reduced IL-13-mediated mitochondrial ROS generation and permeability in MCU knockout tracheal epithelial cells. (A) WT or MCU−/− MTBEC in Transwell inserts were grown to confluence and TEER (ohm × cm 2 ) was measured after cells excluded media from the apical side indicating monolayer establishment. (B) MTBEC were exposed to 10 ng/ml IL-13 for 4 days and epithelial permeability was assessed by the amount of TRITC-dextran (1 mg/ml) detected in the media of the basolateral compartment of the Transwell. Data are represented as fold-change over WT. (C) Quantitation of Mt-ROS with MitoSOX using flow cytometry. Representative plots are shown. All data are the means ± SEM for 3 independent experiments (n = 3 samples/treatment group). Student’s two-tailed t-test was used for (A), one-way ANOVA with Tukey post hoc test for (B, C). * p
Figure Legend Snippet: Reduced IL-13-mediated mitochondrial ROS generation and permeability in MCU knockout tracheal epithelial cells. (A) WT or MCU−/− MTBEC in Transwell inserts were grown to confluence and TEER (ohm × cm 2 ) was measured after cells excluded media from the apical side indicating monolayer establishment. (B) MTBEC were exposed to 10 ng/ml IL-13 for 4 days and epithelial permeability was assessed by the amount of TRITC-dextran (1 mg/ml) detected in the media of the basolateral compartment of the Transwell. Data are represented as fold-change over WT. (C) Quantitation of Mt-ROS with MitoSOX using flow cytometry. Representative plots are shown. All data are the means ± SEM for 3 independent experiments (n = 3 samples/treatment group). Student’s two-tailed t-test was used for (A), one-way ANOVA with Tukey post hoc test for (B, C). * p

Techniques Used: Permeability, Knock-Out, Quantitation Assay, Flow Cytometry, Cytometry, Two Tailed Test

Apoptotic cell death and mitochondrial cytochrome c release by IL-13 are reduced by MCU inhibition. HAEC were infected with Mt-GFP or DN-MCU (10 MOI) for 48 hrs and exposed to IL-13 (10 ng/ml) for 72 hrs. (A) Cell viability by trypan blue staining (n = 5 independent experiments, measured as the ratio of viable cells to total cells). (B) Scatter plots and quantitation of apoptosis based on FITC-Annexin-V/7-AAD staining (n = 5 independent experiments). (C) Representative immunoblots and densitometric analysis for cytochrome C isolated from cytoplasmic fractions of cells (n = 6 independent experiments, results represented as percent of control). (D) Caspase-3 activity was measured via caspase-3 activity assay (n = 4 independent experiments, results are normalized to total protein concentration). All data are the means ± SEM. One-way ANOVA with Tukey post hoc test used. * p
Figure Legend Snippet: Apoptotic cell death and mitochondrial cytochrome c release by IL-13 are reduced by MCU inhibition. HAEC were infected with Mt-GFP or DN-MCU (10 MOI) for 48 hrs and exposed to IL-13 (10 ng/ml) for 72 hrs. (A) Cell viability by trypan blue staining (n = 5 independent experiments, measured as the ratio of viable cells to total cells). (B) Scatter plots and quantitation of apoptosis based on FITC-Annexin-V/7-AAD staining (n = 5 independent experiments). (C) Representative immunoblots and densitometric analysis for cytochrome C isolated from cytoplasmic fractions of cells (n = 6 independent experiments, results represented as percent of control). (D) Caspase-3 activity was measured via caspase-3 activity assay (n = 4 independent experiments, results are normalized to total protein concentration). All data are the means ± SEM. One-way ANOVA with Tukey post hoc test used. * p

Techniques Used: Inhibition, Infection, Staining, Quantitation Assay, Western Blot, Isolation, Activity Assay, Caspase-3 Activity Assay, Protein Concentration

MCU inhibition abolishes ROS production and preserves the membrane potential. (A) Representative images and (B) quantification of mitochondrial ROS production in HAEC infected with Mt-GFP (empty) or DN-MCU adenovirus (10 MOI) prior to exposure to IL-13 (10 ng/ml) for 2 days (n = 5 independent experiments; data were quantified for 5–6 images per treatment at 63x magnification and represented as percent of control). Scale bars = 100μm. (C) Scatter plots and quantification of Mt-ROS of MitoSOX fluoresence by flow cytometry (Representative plots are shown, n = 4 independent experiments, represented as fold change). Representative images of mitochondrial membrane potential staining by TMRM fluorescence. Quantification of TMRM (n=5 independent experiments; data were quantified for 5–6 images per treatment at 63x magnification, represented as percent of control). Scale bars= 100μm. (F) Scatter plots and quantification of TMRM staining by flow cytometry (Representative plots are shown, n = 4 independent experiments, represented as fold change). All data are the means ± SEM. One-way ANOVA with Tukey post hoc test used. * p
Figure Legend Snippet: MCU inhibition abolishes ROS production and preserves the membrane potential. (A) Representative images and (B) quantification of mitochondrial ROS production in HAEC infected with Mt-GFP (empty) or DN-MCU adenovirus (10 MOI) prior to exposure to IL-13 (10 ng/ml) for 2 days (n = 5 independent experiments; data were quantified for 5–6 images per treatment at 63x magnification and represented as percent of control). Scale bars = 100μm. (C) Scatter plots and quantification of Mt-ROS of MitoSOX fluoresence by flow cytometry (Representative plots are shown, n = 4 independent experiments, represented as fold change). Representative images of mitochondrial membrane potential staining by TMRM fluorescence. Quantification of TMRM (n=5 independent experiments; data were quantified for 5–6 images per treatment at 63x magnification, represented as percent of control). Scale bars= 100μm. (F) Scatter plots and quantification of TMRM staining by flow cytometry (Representative plots are shown, n = 4 independent experiments, represented as fold change). All data are the means ± SEM. One-way ANOVA with Tukey post hoc test used. * p

Techniques Used: Inhibition, Infection, Flow Cytometry, Cytometry, Staining, Fluorescence

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    R&D Systems rabbit mabs
    Synergistic and antagonistic effects of multiple <t>NiV-G</t> N-glycans in combination on cell-cell fusion. (A) Relative levels of 293T cell surface expression (HA), binding to <t>MAbs</t> (MAb26, MAb45, and MAb213), binding to soluble ephrinB2 receptor, and cell-cell
    Rabbit Mabs, supplied by R&D Systems, used in various techniques. Bioz Stars score: 94/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems monoclonal anti mouse il 13rα2 antibody
    Human monocytes are more sensitive to IL-4 than to IL-13. (A) Freshly elutriated human monocytes from individual donors were cultured in RPMI containing 2% FBS for 2 h. The cells were left untreated or treated with indicated concentrations of IL-4 or -13 for 15 min. Subsequently, the cells were fixed, permeabilized, and stained with anti-pStat6Y641. Stat6 phosphorylation was measured in the indicated flow cytometry monocyte gate. For each cytokine concentration, MFI of Stat6 phosphorylation was obtained by subtracting the median of the unstimulated sample from the median of the stimulated sample in monocyte gate (bottom). A representative of five independent experiments is illustrated in the flow cytometry plots. The graph relating cytokine concentration to Δ Median represents five independent experiments, with means and SEMs. (B) Elutriated human monocytes were either left unstimulated or were stimulated with the indicated concentrations of cytokines for 30 min at 37°C. Nuclear protein extracts were then prepared, and the levels of Stat6 activity were measured by EMSA using a Stat6 binding site containing radioactively labeled DNA-probe (SBE1). (C) Blocking of γc inhibits IL-4 responses in human monocytes. The experiment was performed as in B, but the cells were pretreated for 1 h with a blocking antibody against human γc. (D) Differential sensitivity of CD16 hi and CD14 hi monocytes toward IL-4 and -13. Elutriated monocytes were treated as in A. The cells were stained with anti-CD14 and -CD16 surface antibodies, before fixing and permeabilizing the cells. After methanol permeabilization, the cells were stained with anti-pStat6Y641 antibody. Staining results from donor number 4 in E are shown. (E) Summary of results of IL-4– and IL-13–induced Stat6 phosphorylation in CD14 hi and CD16 hi human monocytes from four donors; means and SEMs are shown. (F) Expression of IL-4Rα, γc, IL-13Rα1, and <t>IL-13Rα2</t> in CD14 hi and CD16 hi monocyte populations was measured by flow cytometry. The expression was quantitated by subtracting the MFI of specific staining from the MFI of the isotype control and dividing the obtained value by the MFI of the unstained control. Monocytes from seven healthy donors were analyzed; means and SEMs are shown.
    Monoclonal Anti Mouse Il 13rα2 Antibody, supplied by R&D Systems, used in various techniques. Bioz Stars score: 92/100, based on 1 PubMed citations. ZERO BIAS - scores, article reviews, protocol conditions and more
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    R&D Systems il 13
    Th2 cytokines in the lung after sensitization and challenge with PA. Cytokine levels were measured in lung homogenate protein extracts by ELISA on the indicated days after challenge. IL-4 ( A ) and <t>IL-13</t> ( B ) levels in C57BL/6 mice sensitized/challenged
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    Synergistic and antagonistic effects of multiple NiV-G N-glycans in combination on cell-cell fusion. (A) Relative levels of 293T cell surface expression (HA), binding to MAbs (MAb26, MAb45, and MAb213), binding to soluble ephrinB2 receptor, and cell-cell

    Journal: Journal of Virology

    Article Title: N-Glycans on the Nipah Virus Attachment Glycoprotein Modulate Fusion and Viral Entry as They Protect against Antibody Neutralization

    doi: 10.1128/JVI.01304-12

    Figure Lengend Snippet: Synergistic and antagonistic effects of multiple NiV-G N-glycans in combination on cell-cell fusion. (A) Relative levels of 293T cell surface expression (HA), binding to MAbs (MAb26, MAb45, and MAb213), binding to soluble ephrinB2 receptor, and cell-cell

    Article Snippet: The binding of anti-NiV-F antiserum 834 to surface NiV-F and the binding of the anti-NiV-G specific rabbit MAbs (MAb26, MAb45, or MAb213) or mouse anti-hemagglutinin (anti-HA) MAb or soluble ephinB2 fused to human Fc (B2-hFc; R & D Systems, Minneapolis, MN) to cell surface NiV-G were measured by flow cytometry on NiV-F/G-transfected cells.

    Techniques: Expressing, Binding Assay

    Human monocytes are more sensitive to IL-4 than to IL-13. (A) Freshly elutriated human monocytes from individual donors were cultured in RPMI containing 2% FBS for 2 h. The cells were left untreated or treated with indicated concentrations of IL-4 or -13 for 15 min. Subsequently, the cells were fixed, permeabilized, and stained with anti-pStat6Y641. Stat6 phosphorylation was measured in the indicated flow cytometry monocyte gate. For each cytokine concentration, MFI of Stat6 phosphorylation was obtained by subtracting the median of the unstimulated sample from the median of the stimulated sample in monocyte gate (bottom). A representative of five independent experiments is illustrated in the flow cytometry plots. The graph relating cytokine concentration to Δ Median represents five independent experiments, with means and SEMs. (B) Elutriated human monocytes were either left unstimulated or were stimulated with the indicated concentrations of cytokines for 30 min at 37°C. Nuclear protein extracts were then prepared, and the levels of Stat6 activity were measured by EMSA using a Stat6 binding site containing radioactively labeled DNA-probe (SBE1). (C) Blocking of γc inhibits IL-4 responses in human monocytes. The experiment was performed as in B, but the cells were pretreated for 1 h with a blocking antibody against human γc. (D) Differential sensitivity of CD16 hi and CD14 hi monocytes toward IL-4 and -13. Elutriated monocytes were treated as in A. The cells were stained with anti-CD14 and -CD16 surface antibodies, before fixing and permeabilizing the cells. After methanol permeabilization, the cells were stained with anti-pStat6Y641 antibody. Staining results from donor number 4 in E are shown. (E) Summary of results of IL-4– and IL-13–induced Stat6 phosphorylation in CD14 hi and CD16 hi human monocytes from four donors; means and SEMs are shown. (F) Expression of IL-4Rα, γc, IL-13Rα1, and IL-13Rα2 in CD14 hi and CD16 hi monocyte populations was measured by flow cytometry. The expression was quantitated by subtracting the MFI of specific staining from the MFI of the isotype control and dividing the obtained value by the MFI of the unstained control. Monocytes from seven healthy donors were analyzed; means and SEMs are shown.

    Journal: The Journal of Experimental Medicine

    Article Title: Tuning sensitivity to IL-4 and IL-13: differential expression of IL-4R?, IL-13R?1, and ?c regulates relative cytokine sensitivity

    doi: 10.1084/jem.20080452

    Figure Lengend Snippet: Human monocytes are more sensitive to IL-4 than to IL-13. (A) Freshly elutriated human monocytes from individual donors were cultured in RPMI containing 2% FBS for 2 h. The cells were left untreated or treated with indicated concentrations of IL-4 or -13 for 15 min. Subsequently, the cells were fixed, permeabilized, and stained with anti-pStat6Y641. Stat6 phosphorylation was measured in the indicated flow cytometry monocyte gate. For each cytokine concentration, MFI of Stat6 phosphorylation was obtained by subtracting the median of the unstimulated sample from the median of the stimulated sample in monocyte gate (bottom). A representative of five independent experiments is illustrated in the flow cytometry plots. The graph relating cytokine concentration to Δ Median represents five independent experiments, with means and SEMs. (B) Elutriated human monocytes were either left unstimulated or were stimulated with the indicated concentrations of cytokines for 30 min at 37°C. Nuclear protein extracts were then prepared, and the levels of Stat6 activity were measured by EMSA using a Stat6 binding site containing radioactively labeled DNA-probe (SBE1). (C) Blocking of γc inhibits IL-4 responses in human monocytes. The experiment was performed as in B, but the cells were pretreated for 1 h with a blocking antibody against human γc. (D) Differential sensitivity of CD16 hi and CD14 hi monocytes toward IL-4 and -13. Elutriated monocytes were treated as in A. The cells were stained with anti-CD14 and -CD16 surface antibodies, before fixing and permeabilizing the cells. After methanol permeabilization, the cells were stained with anti-pStat6Y641 antibody. Staining results from donor number 4 in E are shown. (E) Summary of results of IL-4– and IL-13–induced Stat6 phosphorylation in CD14 hi and CD16 hi human monocytes from four donors; means and SEMs are shown. (F) Expression of IL-4Rα, γc, IL-13Rα1, and IL-13Rα2 in CD14 hi and CD16 hi monocyte populations was measured by flow cytometry. The expression was quantitated by subtracting the MFI of specific staining from the MFI of the isotype control and dividing the obtained value by the MFI of the unstained control. Monocytes from seven healthy donors were analyzed; means and SEMs are shown.

    Article Snippet: Western blot analysis of murine IL-13Rα2 was performed with monoclonal anti–mouse IL-13Rα2 antibody (R & D Systems).

    Techniques: Cell Culture, Staining, Flow Cytometry, Cytometry, Concentration Assay, Activity Assay, Binding Assay, Labeling, Blocking Assay, Expressing

    Th2 cytokines in the lung after sensitization and challenge with PA. Cytokine levels were measured in lung homogenate protein extracts by ELISA on the indicated days after challenge. IL-4 ( A ) and IL-13 ( B ) levels in C57BL/6 mice sensitized/challenged

    Journal:

    Article Title: Pulmonary Immune Responses to Propionibacterium acnes in C57BL/6 and BALB/c Mice

    doi: 10.1165/rcmb.2005-0285OC

    Figure Lengend Snippet: Th2 cytokines in the lung after sensitization and challenge with PA. Cytokine levels were measured in lung homogenate protein extracts by ELISA on the indicated days after challenge. IL-4 ( A ) and IL-13 ( B ) levels in C57BL/6 mice sensitized/challenged

    Article Snippet: The concentrations of TNF-α, IFN-γ, IL-4, IL-12 p40, IL-12p70, IL-13, and monocyte chemoattractant protein-1 (MCP-1) were measured by ELISA, according to the manufacturer's instructions (R & D Systems Inc., Minneapolis, MN).

    Techniques: Enzyme-linked Immunosorbent Assay, Mouse Assay

    ( A ) Real-time PCR measurements of VEGF mRNA in HASM cells that were left untreated or treated for 6 h with IL-4 (0.3–30 ng/ml) or IL-13 (30 ng/ml). Results are mean ± SE of data from five to nine RNA samples in each case and are normalized

    Journal:

    Article Title: Interleukin-13 and Interleukin-4 Induce Vascular Endothelial Growth Factor Release from Airway Smooth Muscle Cells

    doi: 10.1165/rcmb.2005-0147OC

    Figure Lengend Snippet: ( A ) Real-time PCR measurements of VEGF mRNA in HASM cells that were left untreated or treated for 6 h with IL-4 (0.3–30 ng/ml) or IL-13 (30 ng/ml). Results are mean ± SE of data from five to nine RNA samples in each case and are normalized

    Article Snippet: To examine the importance of +936C/T SNP at the 3′-UTR of the VEGF gene on IL-13–, IL-4–, and TNF-α–induced VEGF release, HASM cells were stratified according to +936 C/T genotype.

    Techniques: Real-time Polymerase Chain Reaction

    Effect of the −460/−152/+405 haplotype on VEGF release from untreated HASM cells (Ctrl) and from cells treated with IL-13 (3 ng/ml), IL-4 (1 ng/ml), or TNF-α (10 ng/ml). Each data point represents the mean value obtained

    Journal:

    Article Title: Interleukin-13 and Interleukin-4 Induce Vascular Endothelial Growth Factor Release from Airway Smooth Muscle Cells

    doi: 10.1165/rcmb.2005-0147OC

    Figure Lengend Snippet: Effect of the −460/−152/+405 haplotype on VEGF release from untreated HASM cells (Ctrl) and from cells treated with IL-13 (3 ng/ml), IL-4 (1 ng/ml), or TNF-α (10 ng/ml). Each data point represents the mean value obtained

    Article Snippet: To examine the importance of +936C/T SNP at the 3′-UTR of the VEGF gene on IL-13–, IL-4–, and TNF-α–induced VEGF release, HASM cells were stratified according to +936 C/T genotype.

    Techniques:

    Effect of +405 genotype ( A ) and of −460/−152 genotype ( B ) on VEGF release under basal conditions (Ctrl) and after treatment with IL-13 (3 ng/ml), IL-4 (1 ng/ml), or TNF-α (10 ng/ml). Each data point represents the mean

    Journal:

    Article Title: Interleukin-13 and Interleukin-4 Induce Vascular Endothelial Growth Factor Release from Airway Smooth Muscle Cells

    doi: 10.1165/rcmb.2005-0147OC

    Figure Lengend Snippet: Effect of +405 genotype ( A ) and of −460/−152 genotype ( B ) on VEGF release under basal conditions (Ctrl) and after treatment with IL-13 (3 ng/ml), IL-4 (1 ng/ml), or TNF-α (10 ng/ml). Each data point represents the mean

    Article Snippet: To examine the importance of +936C/T SNP at the 3′-UTR of the VEGF gene on IL-13–, IL-4–, and TNF-α–induced VEGF release, HASM cells were stratified according to +936 C/T genotype.

    Techniques:

    ( A ) Effects of increasing doses of IL-4 or IL-13 administered for 24 h on VEGF release from HASM cells. Values represent mean ± SE from 4–13 donors. Each donor was studied in duplicate.* P

    Journal:

    Article Title: Interleukin-13 and Interleukin-4 Induce Vascular Endothelial Growth Factor Release from Airway Smooth Muscle Cells

    doi: 10.1165/rcmb.2005-0147OC

    Figure Lengend Snippet: ( A ) Effects of increasing doses of IL-4 or IL-13 administered for 24 h on VEGF release from HASM cells. Values represent mean ± SE from 4–13 donors. Each donor was studied in duplicate.* P

    Article Snippet: To examine the importance of +936C/T SNP at the 3′-UTR of the VEGF gene on IL-13–, IL-4–, and TNF-α–induced VEGF release, HASM cells were stratified according to +936 C/T genotype.

    Techniques:

    Effect of the presence of at least one +936T allele on VEGF release from untreated HASM cells (Ctrl) and from cells treated with IL-13 (3 ng/ml), IL-4 (1 ng/ml), or TNF-α (10 ng/ml). Each data point represents the mean value obtained for

    Journal:

    Article Title: Interleukin-13 and Interleukin-4 Induce Vascular Endothelial Growth Factor Release from Airway Smooth Muscle Cells

    doi: 10.1165/rcmb.2005-0147OC

    Figure Lengend Snippet: Effect of the presence of at least one +936T allele on VEGF release from untreated HASM cells (Ctrl) and from cells treated with IL-13 (3 ng/ml), IL-4 (1 ng/ml), or TNF-α (10 ng/ml). Each data point represents the mean value obtained for

    Article Snippet: To examine the importance of +936C/T SNP at the 3′-UTR of the VEGF gene on IL-13–, IL-4–, and TNF-α–induced VEGF release, HASM cells were stratified according to +936 C/T genotype.

    Techniques: